石油污染物的微生物降解研究

以炼油厂污水池底泥中分离的3株细菌和3株真菌为供试微生物,陕北黄土(0～20 cm)制备的土壤悬浮液为土壤微生物对照,灭菌培养基为非生物降解对照,研究不同组合微生物的生长动态及对石油烃的降解率.将不同组合的微生物接种到石油烃质量浓度为10000 mg·L-1的液体培养基中,29 ℃±1 ℃,摇床连续培养50 d.于0 ～72 h内取样,进行微生物生长动态检测;于5～50 d定期取样,考察培养基中石油烃降解率的动态变化.结果表明:单独外源菌降解效果优于混合外源菌和土著微生物,25 d后土著微生物生物降解率超过混合外源菌.外源细菌、外源真菌、混合外源菌和土著微生物50 d生物降解率分别为71.54 %,60.13 %,47.26 %和51.49 %.土壤细菌对石油污染物具有较强的生长适应性,外源细菌降解效果最好.     

一株选择性降解木质素菌的筛选及其对玉米秸秆的降解

以玉米秸秆为基质,对15株白腐真菌进行了初步筛选,从中获得一株选择性系数较高的菌株Y10,经ITS-5.8SrDNA序列分析,初步鉴定为Cerrena sp.,并研究了该菌在30 d培养期内降解玉米秸秆中木质纤维素的情况.结果表明,菌株Y10对玉米秸秆中木质素和半纤维素的降解速率明显高于纤维素;在30 d的培养过程中,该菌对玉米秸秆降解的选择性系数都大于1,d 15选择性系数最高为3.88.紫外光谱和红外光谱分析结果表明,经菌株Y10降解后玉米秸秆的化学成分发生了变化,且对木质素的降解程度要大于对纤维素的降解程度.图4表3参17     

一株具有高产孢率的头孢霉型虫草菌

编号为CR 95 11的头孢霉型真菌 ,来自一种生长力和出子座率均高的大型新种虫草 ,是该虫草的无性阶段 .对这株真菌在PDA、Czapek’s和Sabouraud’s培养基上的生长性状和形态特征进行了初步的鉴定研究 .连续测定它的动态产孢数量 ,显示CR 95 11具有先缓而后上扬增速极快的产孢趋势 ,比之其它菌株有异常强劲的后续产孢能力和生长力 ,这是菌株对寄主昆虫强侵染力的一个标志特征 .其转折点出现在培养d 7～ 8附近 ,其中在PDA培养d 8的孢子日增加度 (斜率 )达到 2 .7× 10 7,此后加速升高 .将CR 95 11对 2 2株包括细菌、放线菌、丝状真菌和酵母在内的致敏菌株进行抗异实验 ,结果显示 ,CR 95 11只对枯草杆菌、产气杆菌和绿色木霉等 3株显示出明显的抑制能力 ,因而其抗菌活性的规律仍不明确 .然而 ,该菌能与一些酵母类共生 ,生长得到促进 ,是其具备较强生长力和感染力的又一种保证机制 .图 1表 2参 7     

四川攀西干热河谷区根瘤菌的多样性研究

采用数值分类、BOXAIR-PCR和16S rDNA PCR-RFLP方法,对分离自四川攀西干热河谷区11种豆科植物上的43株根瘤菌的表型和遗传多样性进行了研究.数值分类结果表明,攀西地区根瘤菌具有极大的表型性状多样性,在80%相似性水平上绝大多数供试菌株单独成群,能耐60℃(10min处理)的高温,在低温(10℃)或者低pH(4.0)条件下生长很差.与数值分类结果相比较,BOXAIR-PCR的分群结果更分散,说明供试菌株在遗传性状上也具有多样性.在数值分群基础上,选取15个代表菌株进行16S rDNA PCR-RFLP分析.结果表明,供试的15株代表菌株遗传差异明显,仅CCBAU61224与Rhizobium leguminosarum USDA2370T,CCBAU61303与Agrobacterium tumerfacienseIAM13129T具有相同的遗传图谱类型.数值分类和16S rDNA PCR-RFLP分析具有较好的一致性,BOXAIR-PCR适合于对根瘤菌种内菌株间的遗传多样性研究.图4表2参21     

甘薯块根腐烂真菌的分离和鉴定

从腐烂甘薯块根中分离到4株丝状真菌,分别编号为SP-1、SP-2、SP-3和SP-4,它们都能在甘薯块根片上快速生长.将这4株真菌接种在PDA培养基上,观察其菌落形态和结构特征.提取4株菌的总DNA,利用真菌的18S rDNA通用引物进行扩增、克隆和测序,其18S rDNA序列的长度分别为1 810 bp、1 769 bp、1 768 bp和1 770 bp.将这些序列在GenBank中进行序列比对,获得的同源性较高的序列用于构建系统进化树.根据分子鉴定和形态观察的结果,SP-1菌株是匍枝根霉(Rhizopus stolonifer)SP-1,SP-2是尖镰孢(Fusarium oxysporum)SP-2,SP-3是康宁肉座菌(Hypocreakoningii)SP-3,SP-4是肉座菌属(Hypocrea),命名为Hypocrea sp.SP-4.     

一株产木聚糖酶嗜热真菌樟绒枝霉的鉴定及其产纤维质降解酶系分析

对分离得到的一株产耐热木聚糖酶的真菌CAU521进行鉴定,并对其产纤维质降解酶系进行研究.通过菌落形态、显微镜产孢结构以及18S rDNA序列同源性比对等分析,鉴定该菌为樟绒枝霉(Malbranchea cinnamomea),其最适生长温度为45℃,为一株嗜热真菌.该菌能以农业废弃物玉米芯为碳源液体发酵产耐热木聚糖酶,50℃下培养7 d,木聚糖酶的最高酶活力达到173 U/mL.SDS-PAGE和酶谱分析表明该菌株能同时分泌多种纤维质降解酶:4种木聚糖酶、2种纤维素酶、3种葡聚糖酶和1种甘露聚糖酶.结果表明樟绒枝霉CAU521在降解和利用纤维质材料方面具有潜在的应用价值.     

一种白腐真菌的分离、鉴定、培养及产漆酶条件

白腐真菌是一类降解木质素使木材形成白色腐朽的担子菌,往往具有高产胞外漆酶的活性.从北京农学院校园内分离到一株高温型、高产漆酶白腐真菌菌株,编号为BUA-01.结合形态学特征与ITS序列分析,确定其分类学地位;进一步研究其菌丝生长的最适碳源、氮源、碳氮比(C/N)、生长因子、最适温度和最适p H值;利用不同浓度Cu~(2+)(Cu SO4)的诱导,探讨液体发酵条件下对胞外漆酶产量的影响;通过向发酵液中加入3种偶氮染料(依文思蓝、甲基橙和铬黑T),研究菌株对染料的降解效果.结果表明,该菌株与毛栓菌(Trametes hirsuta)的同源性最高,为100%,遗传距离为0,确定BUA-01菌株为栓菌属(Trametes)真菌.菌丝体生长的最适碳源为淀粉,最适氮源为黄豆粉、酵母浸粉,最适C/N值为40/1和10/1,最适温度为37℃,最适p H为6.0-7.0,供试生长因子对菌丝生长无显著促进作用.0.25m mol/L的Cu~(2+)对胞外漆酶产量有极显著的促进作用,在96 h时,发酵液的活性达到最高,为1081.33±6.3 U/m L,是对照组的26倍.BUA-01菌株对偶氮染料降解效果显著,12 h对依文思蓝、甲基橙和铬黑T的脱色率分别为93.31%±0.16%、92.37%±0.42%和79.25%±0.64%.本研究表明菌株BUA-01在产胞外漆酶和染料降解方面具有潜在应用价值.     

土壤中降解聚己内酯(PCL)微生物菌种的筛选及降解性能

为了获得土壤中能降解聚己内酯(PCL)的微生物并研究其降解效果,对陕西普通土壤中的细菌、真菌、霉菌和放线菌进行驯化、筛选,并分别对PCL进行生物降解,研究降解前后PCL质量损失、表面形态、结晶度三方面的变化,结果表明,在陕西普通土壤中存在能够降解PCL的细菌、真菌、放线菌,其中放线菌的降解能力最好,真菌中霉菌的降解能力相对较弱,部分细菌具有较好的降解能力.经生理生化实验鉴定:具有较强降解PCL的细菌为蜡状芽孢杆菌.经上述菌种侵蚀后的PCL球晶均受到一定程度的破坏,其结晶度都有所降低.     

多效唑在土壤中降解、吸附和淋溶作用

本文研究了植物生长调节剂多效唑(MET)在土壤中的环境引为,MET在土壤中降解较缓慢并随土壤环境条件而异,淹水厌氧条件比旱地好氧条件降解更缓慢,灭菌土壤中未见明显降解,微生物培养试验表明,MET对真菌有明显抑止作用,以MET为唯一营养源的培养基上能生长出几种放线菌,在三种供试土壤中,MET在高有机质含量的黑土中吸附常数远大于黄棕壤和潮土;MET的淋溶则在砂粒含量较高和有机质含量较低的潮土中最快,黑土最慢,黄棕壤居间.     

盐京九号水稻秸秆对铜绿微囊藻（Microcystis aeruginosa）的抑制作用

以盐京九号水稻为材料,研究经高温灭菌或物理粉碎后的水稻秸秆微细颗粒对铜绿微囊藻(Microcystisaeruginosa)生长的抑制及其作用机制.结果表明,当藻液中水稻秸秆添加量≥10g·L-1时,高温灭菌与未灭菌的水稻秸秆同样能抑制微囊藻生长.灭菌稻秆试验组藻细胞的呼吸速率和SOD活性都呈现出先上升后迅速下降的趋势;通过有机溶剂(石油醚、乙酸乙酯和乙醚)萃取稻秆水浸液,用滤纸片法在固体培养基上所做的抑藻试验结果表明,乙醚提取液有明显的抑藻圈.提示抑藻效应是水稻秸秆中某些化学物质溶出而产生的作用,与微生物的活动无关.进一步分离鉴定乙醚提取液结果显示,浸提液中有机酸类和酚类物质有显著的抑藻效应.水稻秸秆微粒的粒径越小,产生的抑藻效应出现得越早,也越明显,这可能还与秸秆微粒的吸附作用有关.     

菌根真菌对土壤呼吸的影响

土壤是陆地生态系统的重要组成部分,是地球最大的碳库之一。土壤呼吸是陆地生态系统向大气释放CO2的主要途径之一,其微小的变化将导致大气CO2浓度的较大波动。菌根是土壤真菌与植物根系形成的共生体,存在于绝大多数植物(90%)的根系和生境中。菌根共有7种类型,其中,在自然界中以丛枝菌根和外生菌根为主。众多研究表明,菌根对土壤呼吸有着至关重要的影响,是预测土壤CO2释放速率必须考虑,但却是难以估算的因素。文章总结了有关菌根(包括丛枝菌根和外生菌根)对土壤呼吸影响的研究进展,对目前所得到的研究结果进行了分析,表明菌根真菌侵染植物根系形成菌根后,能提高土壤呼吸的速率,其可能的途径有3条:(1)增强了根系的呼吸,(2)菌根真菌自身呼吸的组分,(3)根外菌丝促进了非根际区土体的呼吸。但是,菌根侵染对根系呼吸敏感性(Q10)影响的研究,大多数则表现为不显著。同时,菌根对土壤呼吸的影响受到各种因素的制约。通过对不同温度下菌根真菌呼吸速率的分析,表明菌根真菌对温度的升高具有适应性。从目前已发表的报道来看,目前关于菌根对土壤呼吸影响的研究还非常少,但可喜的是,近年来,越来越多的研究已经意识到了菌根在土壤呼吸中的重要作用。准确评估菌根在土壤呼吸中的贡献,将有助于预测未来在气候变化下,土壤CO2的排放量。     

菌根真菌对土壤呼吸的影响

土壤是陆地生态系统的重要组成部分,是地球最大的碳库之一。土壤呼吸是陆地生态系统向大气释放CO2的主要途径之一,其微小的变化将导致大气CO2浓度的较大波动。菌根是土壤真菌与植物根系形成的共生体,存在于绝大多数植物（90％）的根系和生境中。菌根共有7种类型,其中,在自然界中以丛枝菌根和外生菌根为主。众多研究表明,菌根对土壤呼吸有着至关重要的影响,是预测土壤CO2释放速率必须考虑,但却是难以估算的因素。文章总结了有关菌根（包括丛枝菌根和外生菌根）对土壤呼吸影响的研究进展,对目前所得到的研究结果进行了分析,表明菌根真菌侵染植物根系形成菌根后,能提高土壤呼吸的速率,其可能的途径有3条：（1）增强了根系的呼吸,（2）菌根真菌自身呼吸的组分,（3）根外菌丝促进了非根际区土体的呼吸。但是,菌根侵染对根系呼吸敏感性（Q10）影响的研究,大多数则表现为不显著。同时,菌根对土壤呼吸的影响受到各种因素的制约。通过对不同温度下菌根真菌呼吸速率的分析,表明菌根真菌对温度的升高具有适应性。从目前已发表的报道来看,目前关于菌根对土壤呼吸影响的研究还非常少,但可喜的是,近年来,越来越多的研究已经意识到了菌根在土壤呼吸中的重要作用。准确评估菌根在土壤呼吸中的贡献,将有助于预测未来在气候变化下,土壤cO2的排放量。     

Effects of Arbuscular mycorrhizal fungi isolated from white clovers (Trifolium repens L) on soil bacteria and fungi

White clover potted experiments were performed to investigate the effects of seven indigenous arbuscular mycorrhizal fungi (AMF) communities isolated from different test plots subjected to long-term fertilisation on soil enzyme activities, number of soil bacteria and fungi. The results showed that the inoculation of arbuscular mycorrhizal fungi communities increased the mycorrhizal infection rate of the plants and promoted the growth of plants. The Mnp treatment was most effective. The shoot biomass, root biomass, potassium and nitrogen uptake of the white clover in Mnp treatment group were increased by 61.54%, 84.00%, 62.50% and 46.71% respectively, compared with those in non-inoculation treatment. The inoculation of AMF communities had little effect on the number of bacteria in the soil, but significantly increased the number of soil fungi. Mnk treatment group had the highest number of fungi in the soil, which was 9.91 times that of the non-inoculation treatment group. The catalase and dehydrogenase activities were both significantly improved in Mnp treatment by 28.12% and 205.38% respectively, compared with those of the control treatment (-M). The urease, invertase and cellulase activities reached the highest levels in the Mck treatment; they were increased by 142.79%, 41.17% and 77.62% respectively, compared with those of the control treatment. Pearson correlation analysis showed that the soil enzyme activity was not correlated with the mycorrhizal infection rate, but correlated with the spore number of the AMF community. The impact of AMF community on soil quality is important for us to understand the function of the ecosystems. Relevant study provides important guidance for maintaining the balance of the soil-plant system and the development of sustainable agriculture.     

Effect of the endectocide ivermectin on filamentous fungi

We investigated the effect of ivermectin, at environmental concentrations and above, on several strains of filamentous fungi. Ivermectin did not show significant short-term effect on biomass production and germination rate of the spores. By contrast, the spore production was stimulated twofold in Phanerochaete chrysosporium and Mucor racemosus cultures, whereas that endpoint was reduced in Fusarium cultures. A long-term exposure of F. oxysporum to ivermectin reduced both production and germination of fungal spores. Ivermectin was not an inducer of exocellular oxidases.     

Antimicrobial activities of flavonoids against bamboo-destroying fungi and molds

It is well-known that Poria vaporaria (Pers.:Fr.) Cooke, Paecilomyces variotii Bainier and Aspergillus niger are fungi and molds which destroy bamboo. The aim of this study was to compare the effectiveness of antibacterial activity of 16 flavonoids in preventing the actions of these three bamboo-destroying fungi and molds (Poria vaporaria (Pers.:Fr.) Cooke, Paecilomyces variotii Bainier and Aspergillus niger). The results showed that baicalein(1) exerted the highest inhibitory effect on Poria vaporaria (Pers.:Fr.) Cooke and Paecilomyces variotii Bainier indicating that baicalein(1) may be considered as a potential agent to develop as natural bactericide to preserve natural bamboo.     

Marine fungi deteriorating chitin of hydrozoa and keratin-like annelid tubes

Higher marine fungi were found for the first time as degraders of chitinous exoskeletons of hydrozoa and of keratinous (?) tubes of annelids. Collections were made in several locations of the Atlantic and Pacific Oceans. Fruiting bodies and hyphae of the ascomycete Abyssomyces hydrozoicus Kohlm. occurred on hydrozoa at a depth of 631 to 641 m near the South Orkney Islands. Sterile mycelia, some of them resembling Dictyonema zoophytarum Reinsch, were collected on hydrozoa on the North Carolina coast, and offshore at a depth of 46 to 73 m. Tubes of the polychaete Chaetopterus variopedatus (Renier) washed ashore in California contained ascocarps and mycelia of Lulworthia sp. Hyphal growth on and in the animal tubes is described and illustrated.     

丛枝菌根真菌对羊草凋落物降解作用的研究

丛枝菌根真菌（Abuscular mycorrhizal fungi）能够影响植物生长及养分含量,从而影响凋落物的降解。采用根袋的方法研究了接种两种丛枝真菌Glomus mosseae和Glomus claroideum对羊草（Leymus chinensis）地上部及根系凋落物降解的影响。结果表明,随时间的延长,凋落物的重量逐渐减少,凋落物氮、磷含量均表现出先下降后逐渐升高的趋势。接种对地上部凋落物的养分含量及降解速度未产生显著性影响,但显著降低了根系氮磷含量及降解系数。接种Glomus mosseae和Glomus claroideum羊草根系氮、磷含量均显著低于CK;接种与未接种相比羊草根系k值显著降低;根系C：N未接种处理显著低于接种处理。说明丛枝菌根真菌可能间接影响草原生态系统中有机物质的分解和养分释放。     

菌根真菌增加植物抗盐碱胁迫的机理

菌根是菌根真菌与高等植物根系形成的一种共生体，能够促进植物在瘠薄土壤中的生长。文章通过综合近20年来国内外在菌根植物抗盐碱研究方面的成就，论述了在高盐胁迫下，菌根对其寄主植物耐盐碱能力的影响；指出菌根植物可能主要通过以下4个方面增加对高浓度盐碱环境的抗性：增加植株对P等矿质营养元素的吸收，改善了盐胁迫引起的营养亏缺；改变植物体内离子平衡，降低其生理毒害；增加植株对水分的吸收和利用能力，缓解了生理性干旱；改变了植物根系形态，促进根系水分吸收能力。在分析菌根真菌增加植物抗盐碱胁迫机理的基础上，还对该问题的研究前景提出了设想，为盐碱地改良提供参考依据。     

Incidence of keratinophilic fungi in waterlogged condition of paddy soil

Prevalence of keratinophilic fungi was observed in paddy field soil during different stages of cultivation viz., transplanting, tillering, milking and maturation. Out of total 76 soil samples, 65 soil samples were found to be positive for the keratinophilic fungi. Fourteen species belonging to a single genus Chrysosporium were isolated through out the cropping season. C. keratinophilum (17.1%) followed by C. tropicum (13.15%) were found to be the most dominating geophilic species. The highest percent distribution of keratinophilic fungi appeared during the milking stage (100%) of paddy cultivation, followed by the maturation stage (89.47%).     

Application of next-generation sequencing technologies to conservation of wood-inhabiting fungi

Next-generation sequencing (NGS) has significantly increased knowledge of microbial communities and their distribution. However, it is still not common to apply NGS technology to microbial conservation. We sought to use NGS technologies to evaluate conservation strategies for wood-inhabiting fungi. Evaluating a deadwood experiment 3 years after it was established, we specifically examined which tree species combinations promoted the highest richness of wood-inhabiting fungi. Deadwood enrichment was an effective strategy and logs of 6 tree species, either those with the highest wood-inhabiting fungal α and γ diversity or those with the highest β diversity, maintained >1,000 operational taxonomic units (OTUs) spread over a wide range of taxonomic groups. In comparison, a conservation strategy based only on the results of sporocarp surveys yielded 591 OTUs. This result highlights the need to use NGS approaches to inform microbial conservation strategies. We also determined that 5 tree species with the highest saproxylic beetle γ diversity simultaneously conserved wood-inhabiting fungi. Apart from deadwood volume, we suggest data on deadwood quality and species also be included as indicators, especially for wood-inhabiting fungal diversity, and incorporated quickly in forest assessment and monitoring systems in Central Europe.